基于UPLC-Q-TOF-MS分析通络清脑注射剂在大鼠体内的代谢产物

目的:研究大鼠尾静脉注射通络清脑注射剂后尿液中的代谢产物,为该制剂的作用机制分析提供参考。方法:大鼠尾静脉注射通络清脑注射剂,收集0~12 h尿液,采用超高效液相色谱-四级杆-飞行时间串联质谱(UPLC-Q-TOF-MS)分析尿液样品,流动相0.1%甲酸水溶液-乙腈梯度洗脱,流速0.3 m L·min~(-1),电喷雾离子源,正离子模式,扫描范围m/z 100~1 400。结果:检测并鉴定到7个原型成分(黄芩苷,栀子苷,三七皂苷R1,人参皂苷Rg1,Rb1,Rd和Re)及10个可能的代谢产物(其中4个来源于黄芩苷,6个来源于栀子苷)。10个可能的代谢产物分别为葡萄糖基化代谢物(B-M1,G-M2和G-M4),葡萄糖醛酸化代谢物(B-M2),甲基化代谢物(B-M3),水解代谢物(B-M4),去甲基化代谢物(G-M1),脱氢代谢物(G-M3)和羟基化代谢物(G-M5和G-M6),而人参皂苷类成分则暂时未能在尿液中检测到代谢产物。结论:尾静脉注射通络清脑注射剂后,大鼠尿液中能检测到黄芩苷和栀子苷的原型及不同途径的10个代谢产物,而三七总皂苷各成分则只检测到药物原型,暂时未能检测到代谢产物。

Analysis on in Vivo Metabolites of Tongluo Qingnao Injection in Rats by UPLC-Q-TOF-MS

Objective: To investigate the metabolites of Tongluo Qingnao injection in urine of rats after tail vein administration. Method: An UPLC-Q-TOF-MS method was used to analysis the metabolites of 12 h urine samples of rats after tail vein administration of Tongluo Qingnao injection. Result: Seven prototype components (baicalin,geniposide,notoginsenoside R₁,ginsenoside Rg₁,Rb₁,Rd,Re) and 10 metabolites (4 from baicalin and 6 from geniposide) were detected and identified in rat urine,including glycosylated metabolites (B-M1,G-M2 and G-M4),glucuronide metabolite (B-M2),methylated metabolite (B-M3),hydrolytic metabolite (B-M4),demethylation metabolite (G-M1),dehydrogenation metabolite (G-M3) and hydroxylated metabolites (G-M5 and G-M6),while the ginsenosides were temporarily unable to detect metabolites in the urine. Conclusion: Prototype components and metabolites of multiple metabolic pathways of baicalin and geniposide are detected in rat urine after tail vein administration of Tongluo Qingnao injection,but notoginseng total saponins are only detected in the prototype and temporarily failed to detect metabolites.